RNA-seq of dogs: young adult whole blood
Three ml blood was taken from the vena cephalica or the vena saphena lateralis in accordance with the principles of lege artis by an experienced veterinarian in the presence of the dogs owner. In order to preserve the RNA fraction of the blood as intact as possible, it was collected directly into DNA/RNABlood Collection Tubes (Zymo Research). Blood was gently mixed with the special RNA preservative fluid and stored in -20 C. Quick-DNA/RNA Blood Tube kits (Zymo Research) were used for RNA extraction. Isolated RNA was stored in -80 C ultra-low temperature freezer until further processing.RNA samples were treated with DNase in order to remove DNA contamination from the RNA samples. Protein coding RNA was purified via poly(A) capture, and library preparation was performed with the TruSeq Stranded mRNA Library preparation kit (Illumina, CA, USA). Sample quality control was implemented after the DNase treatment and after the library preparation alike (RIN: 8.5-10.0 in all samples).